Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

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Schilsky, M.L. et al.

Schilsky, Stockert, Kesner, Gorla, Gagliardi, Terada, Miura, Czaja,

Copper resistant human hepatoblastoma mutant cell lines without metallothionein induction overexpress ATP7B.

Mutant human hepatoblastoma cell lines resistant to copper toxicity were isolated from mutagenized HuH7. Two copper resistant cell lines (CuR), CuR 23 and CuR 27, had reduced basal expression of metallothionein (MT) messenger RNA (mRNA) and exhibited minimal or no increase in resistance to cadmium or zinc toxicity. Copper uptake, efflux of newly transported copper, glutathione content, and efflux rate were comparable with HuH7, whereas holoceruloplasmin synthesis and secretion were slightly decreased. Subcellular distribution of copper at steady-state showed an increase in organelle and membrane fractions with a reduction in cytosol. Expression of ATP7B mRNA was fivefold increased, and ATP7B protein approximately threefold increased in both CuR 23 and 27. Another cell line, CuR 41, showed increased basal expression of MT and ATP7B mRNA but not ATP7B protein, and resistance to cadmium and zinc toxicity. Copper uptake in CuR 41 was comparable with HuH7, but initial rates of efflux of copper and glutathione were reduced. The synthesis of holoceruloplasmin but not ceruloplasmin peptide was markedly diminished in CuR 41. Subcellular distribution of copper showed an increase in cytosolic and decreased organelle and membrane-associated copper. These data suggest that cellular resistance to copper toxicity was achieved in two independent cell lines without MT induction and that the induction of ATP7B may lead to the enhanced intracellular sequestration of copper by organelles.[1]

References

Copper resistant human hepatoblastoma mutant cell lines without metallothionein induction overexpress ATP7B. Schilsky, M.L., Stockert, R.J., Kesner, A., Gorla, G.R., Gagliardi, G.S., Terada, K., Miura, N., Czaja, M.J. Hepatology (1998) [Pubmed]

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